Filling device and filling method

ABSTRACT

A filling method is provided. The filling method has a filling step in which a valve at the tip of a vertically movable nozzle of a filling device is disposed within a container and a liquid containing at least either or both of cells of a microorganism and a substance treated thereby transferred from a storage tank is sprayed from the valve. In the filling step, either or both of an initial flow rate for filling and a later stage flow rate for filling are less than the maximum flow rate. In addition, the initial filling stage in this filling step, the valve at the tip of the movable nozzle is disposed in the proximity of the bottom surface of the container, and thereafter, the movable nozzle is made to rise according to the filling conditions for the liquid in the container.

TECHNICAL FIELD

The present invention relates to a filling device and a filling method for filling a liquid containing bacterial cells of a microorganism and/or a treatment substance obtained therefrom stored in a storage tank into a container.

The present application claims priority based on Japanese Patent Application No. 2011-196036 filed on Sep. 8, 2011, of which the disclosure is incorporated herein by reference.

BACKGROUND ART

In recent years, a trend of using a biocatalyst became more and more intensive from the viewpoint of effectively using biochemical reaction having features such as energy-saving, resource-saving and low-pollution process in production of a useful substance, and the like. As the biocatalyst, used are animal cells, plant cells, cell organelles, bacterial cells of a microorganism, a treatment substance obtained therefrom and the like containing enzymes that catalyze the intended reaction. Examples of such enzyme include nitrile hydratases having hydration ability for a nitrile group, nitrilases having hydrolysis ability for a nitrile group, or amidases having hydrolysis ability for an amide group, and the like. Use of these enzymes allows cheap manufacture of amides, carboxylic acids, and the like that are important in the chemical industry.

In recent years, not only a reaction pattern of the enzyme reaction, but also peripheral technologies such as a storage method and filling method for a biocatalyst were improved or suggested from spread of use of biocatalysts in the field of the industrial process. For example, in Patent Document 1, it is suggested a method of storing a liquid containing a microorganism, for example, a microorganism containing a nitrile hydratase, bacterial cells or a treatment substance obtained therefrom before subjecting the liquid to thermal disinfection, in which the microorganism is aerated and stirred with the pH of the liquid adjusted to 6 to 8.

REFERENCE LIST Patent Document

Patent Document 1: Japanese Patent No. 4205332

DISCLOSURE OF THE INVENTION Problem to be Solved by the Invention

A liquid containing bacterial cells of a microorganism and/or a treatment substance obtained therefrom easily foams. For that reason, there are problems that the liquid foams when the liquid is filled into a container from a tank in which the liquid is stored, and the filling rate per volume of the container decreases.

The invention has been accomplished under the circumstances described above, and the object thereof is to provide a filling device and a filling method that make it possible to effectively fill a liquid containing bacterial cells of a microorganism and/or a treatment substance obtained therefrom into a container.

Means for Solving Problem

The invention to resolve the problems has the aspects described below.

[1] A filling device for filling a liquid containing bacterial cells of a microorganism and/or a treatment substance obtained therefrom stored in a storage tank into a container,

which is equipped with a nozzle that is disposed in a container, and discharges the liquid transferred from the storage tank into the container, wherein the liquid discharge section of the nozzle is disposed in the filling liquid in the middle of the filling process of the liquid.

[2] A filling device for filling a liquid containing bacterial cells of a microorganism and/or a treatment substance obtained therefrom stored in a storage tank into a container,

which is equipped with a nozzle that discharges the liquid transferred from the storage tank into the container; a detection means that detects the status of the filling of the liquid in the container; and a flow rate-controlling means that controls the flow rate of the liquid discharged from the nozzle depending on the status of the filling of the liquid in the container.

[3] A filling device for filling a liquid containing bacterial cells of a microorganism and/or a treatment substance obtained therefrom stored in a storage tank into a container,

which is equipped with a vertically movable nozzle that discharges the liquid transferred from the storage tank into the container; a detection means that detects the status of the filling of the liquid in the container; and a nozzle position-controlling means that controls the position of the tip of the movable nozzle depending on the status of the filling of the liquid in the container.

[4] The filling device according to any one of claims 1 to 3, which is constituted such that the storage tank is positioned upward from the filling device, and the liquid is filled into the container by the gravity.

[5] The filling device according to any one of claims 1 to 4, wherein the bacterial cells of a microorganism and/or a treatment substance obtained therefrom have nitrile hydratase activity.

[6] A filling method for filling a liquid containing bacterial cells of a microorganism and/or a treatment substance obtained therefrom stored in a storage tank into a container, the filling method including a filling step in which the tip of a nozzle of a filling device is positioned in the container, and the liquid is discharged from the tip, wherein the filling device is equipped with

a nozzle that discharges the liquid transferred from the storage tank into the container; a detection means that detects the status of the filling of the liquid in the container; and a flow rate-controlling means that controls the flow rate of the liquid discharged from the nozzle depending on the status of the filling of the liquid in the container, and wherein

in the filling step, the flow rate at the initial stage for filling and/or the flow rate at the later stage for filling are less than the maximum flow rate.

[7] The filling method according to claim 6, wherein the flow rate at the initial stage for filling of the filling step is equal to or less than a half of the maximum flow rate.

[8] The filling method according to claim 6 or 7, wherein the flow rate at the later stage for filling of the filling step is equal to or less than a half of the maximum flow rate.

[9] A filling method for filling a liquid containing bacterial cells of a microorganism and/or a treatment substance obtained therefrom stored in a storage tank into a container, the filling method including a filling step in which the tip of a movable nozzle of a filling device is positioned in the container, and the liquid is discharged from the tip, wherein the filling device is equipped with

a vertically movable nozzle that discharges the liquid transferred from the storage tank into the container; a detection means that detects the status of the filling of the liquid in the container; and a nozzle position-controlling means that controls the position of the tip of the movable nozzle depending on the status of the filling of the liquid in the container, and wherein

the tip of the movable nozzle is positioned in the proximity of the bottom surface of the container at the initial stage for filling of the filling step, and thereafter, the movable nozzle is made to rise depending on the status of the filling of the liquid in the container.

[10] The filling method according to any one of claims 6 to 9, wherein the bacterial cells of a microorganism and/or the treatment substance obtained therefrom have nitrile hydratase activity.

Specifically, the invention relates to the followings.

(1) A filling device for filling a liquid containing at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom stored in a storage tank into a container, which is equipped with a nozzle that is disposed in a container, and discharges the liquid transferred from the storage tank into the container, wherein the liquid discharge section of the nozzle is disposed in the filling liquid in the middle of the filling process of the liquid.

(2) A filling device for filling a liquid containing at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom stored in a storage tank into a container, which is equipped with a nozzle that discharges the liquid transferred from the storage tank into the container; a detection means that detects the status of the filling of the liquid in the container; and a flow rate-controlling means that controls the flow rate of the liquid discharged from the nozzle depending on the status of the filling of the liquid in the container.

(3) A filling device for filling a liquid containing at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom stored in a storage tank into a container, which is equipped with a vertically movable nozzle that discharges the liquid transferred from the storage tank into the container; a detection means that detects the status of the filling of the liquid in the container; and a nozzle position-controlling means that controls the position of the tip of the movable nozzle depending on the status of the filling of the liquid in the container.

(4) The filling device according to any one of (1) to (3), which is constituted such that the storage tank is positioned upward from the filling device, and the liquid is filled into the container by the gravity.

(5) The filling device according to any one of (1) to (4), wherein the at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom have nitrile hydratase activity.

(6) A filling method for filling a liquid containing at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom stored in a storage tank into a container, the filling method including a filling step in which the tip of a nozzle of a filling device is positioned in the container, and the liquid is discharged from the tip, wherein the filling device is equipped with a nozzle that discharges the liquid transferred from the storage tank into the container; a detection means that detects the status of the filling of the liquid in the container; and a flow rate-controlling means that controls the flow rate of the liquid discharged from the nozzle depending on the status of the filling of the liquid in the container, and wherein in the filling step, at least either or both of the flow rate at the initial stage for filling and the flow rate at the later stage for filling are less than the maximum flow rate.

(7) The filling method according to (6), wherein the flow rate at the initial stage for filling of the filling step is equal to or less than a half of the maximum flow rate.

(8) The filling method according to (6) or (7), wherein the flow rate at the later stage for filling of the filling step is equal to or less than a half of the maximum flow rate.

(9) A filling method for filling a liquid containing at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom stored in a storage tank into a container, the filling method including a filling step in which the tip of a movable nozzle of a filling device is positioned in the container, and the liquid is discharged from the tip, wherein the filling device is equipped with a vertically movable nozzle that discharges the liquid transferred from the storage tank into the container; a detection means that detects the status of the filling of the liquid in the container; and a nozzle position-controlling means that controls the position of the tip of the movable nozzle depending on the status of the filling of the liquid in the container, and wherein the tip of the movable nozzle is positioned in the proximity of the bottom surface of the container at the initial stage for filling of the filling step, and thereafter, the movable nozzle is made to rise depending on the status of the filling of the liquid in the container.

(10) The filling method according to any one of (6) to (9), wherein the at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom have nitrile hydratase activity.

Effect of the Invention

According to the invention, it is possible to provide a filling device and a filling method, which make it possible to effectively fill a liquid containing at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom into a container.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 is a schematic drawing that illustrates the constitution of a filling device 10 used in Example 1.

BEST MODE(S) FOR CARRYING OUT THE INVENTION

The filling device and the filling method of the invention are a filling device and filling method for filling a liquid containing at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom stored in a storage tank into a container.

Herein, the bacterial cells of a microorganism used in the invention may be live bacterial cells, or killed bacterial cells.

Examples of the treatment substance obtained from the bacterial cells include substances obtained by crushing bacterial cells (live bacterial cells or killed bacterial cells), extracts from bacterial cells, immobilized substances obtained by immobilization of bacterial cells or extracts thereof on a carrier, and the like. The extracts are preferably enzymes (coarse enzymes or purified enzymes).

Examples of the method for immobilizing the bacterial cells or extracts thereof include entrapment method, crosslinking method, or carrier-binding method and the like. The entrapment method is a method in which bacterial cells or enzymes are incorporated into minute lattice of a polymer gel, or coated by a film of a semipermeable polymer. The crosslinking method is a method in which enzymes are crosslinked with a reagent having two or more functional groups (multifunctional crosslinking agent). The carrier-binding method is a method in which enzymes are bonded to a water-insoluble carrier.

Examples of the carrier used in the immobilization include, for example, glass beads, silica gel, polyurethane, polyacrylic amide, polyvinyl alcohol, carrageenan, alginic acid, agar, or gelatin and the like.

The at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom are preferably those having enzyme activity that allows the bacterial cells and the treatment substance to be used as a biocatalyst. Examples of the enzyme activity include, for example, nitrile hydratase activity, nitrilase activity, or amidase activity and the like. Among these, the enzyme activity is preferably nitrile hydratase activity.

The nitrile hydratase activity means an activity acting on a nitrile group of a nitrile compound whereby to convert the nitrile compound to a corresponding amide compound. Examples of a method for measuring the nitrile hydratase activity include, for example, a method in which the reaction speed per 1 mg of dry bacterial cells is measured when reacted at a reaction temperature of 10° C. for 10 minutes, and the like.

50 to 500 U/mg of at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom having nitrile hydratase activity is preferably used in the filling method of the invention for filling a liquid containing at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom stored in a storage tank into a container.

Examples of the at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom having nitrile hydratase activity include bacterial cells of a microorganism containing a nitrile hydratase, nitrile hydratase extracted from the bacterial cells, immobilized substances obtained by immobilization of the bacterial cells or nitrile hydratase, and the like.

As the microorganism containing a nitrile hydratase, any microorganism may be used if it is a microorganism having catalytic activity of converting a nitrile to an amide (nitrile hydratase activity). Examples of the microorganism containing a nitrile hydratase may include, for example, microorganisms belonging to Bacillus genus, Bacterinium genus, Micrococcus genus, Brevibacterium genus, Corynebacterium genus, Nocardia genus, Pseudomonas genus, Microbacterium genus, Rhodococcus genus, Gordona genus, Vibrio genus, Nitrosomonas genus, Streptococcus genus, Lactobacillus genus, Attotobacter genus, Saccharomyces genus, Endomyces genus, Aspergillus genus, Penicillium genus, Mucor genus, Rhizopus genus, Achromobacter genus, Pscudonocardia genus, Fusarium genus or Agrobacterium genus, and the like. The microorganism containing a nitrile hydratase is preferably microorganisms belonging to Rhodococcus genus.

In addition, a transformant may be used, which is obtained by acquiring a target gene such as nitrile hydratase, nitrilase or amidase derived from the microorganism, and introducing the gene as it is, or as artificially improved into any host by a conventional method (see Molecular Cloning 2nd Edition, Cold Spring Harbor Laboratory Press, 1989). Examples of such transformant may include, for example, Escherichia coli MT10770 transformed with nitrile hydratase of a bacterium of the Achromobacter genus (FERM P-14756) (see Japanese Patent Application Laid-Open (JP-A) No. 8-266277), Escherichia coli MT10822 transformed with nitrile hydratase of a bacterium of the Pseudonocardia genus (FERM BP-5785) (see JP-A No. 9-275978), or a microorganism transformed with nitrile hydratase of a species of Rhodococcus rhodochrous (see JP-A No. 4-211379), and the like.

The microorganism may be used alone of any one kind, or may be used in combination of 2 or more kinds.

Examples of the liquid containing at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom include a culture liquid obtained by culturing a microorganism in a liquid culture medium; a suspension of resting bacterial cells obtained by isolation from the culture liquid, and washing if desired; or a bacterial cell-crushed liquid obtained by crushing the suspension of resting bacterial cells with ultrasonic wave or metal beads and the like under low temperature; and the like.

As the culture conditions (composition or pH of a liquid culture medium, or culture temperature, and the like), known culture conditions may be adopted depending on a microorganism to be cultured. For example, when the microorganism is a microorganism having nitrile hydratase activity, the microorganism is cultured on a liquid culture medium added with a carbon source (saccharides such as glucose or fructose), a nitrogen source (an inorganic nitrogen source such as ammonium sulfate, ammonium chloride or ammonium nitrate, or an organic nitrogen source such as yeast extract, peptone or meat extract) and if desired, inorganic salts, metal salts, or vitamin and the like at 20 to 40° C. and pH 5 to 9. The culture may be suitably a shaking culture or stirring culture.

Thus-obtained liquid containing at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom is usually stored in a storage tank, and thereafter, the liquid is filled into a container for transport or storage.

The storage tank stores the liquid during the time between the manufacture and the filling into a container, and thus a storage tank that is made of a metal and can control the temperature of the liquid to be stored to a temperature suitable for the storage (for example, equal to or less than 5° C.) is suitably used industrially as the storage tank.

The container is not particularly limited if it is a container appropriate for filling a liquid, and a container that has been used so far for transporting or storing a liquid containing at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom may be used. Generally, a drum or tank made of a metal or made of a resin, and the like may be used as the container. As the drum, those having an internal volume of 200 L or so are usually used. As the tank, those having an internal volume of 1 to 10 m³ or so are usually used.

The filling device and the filling method of the invention are used when filling the liquid from this storage tank into the container.

The filling device of the invention is equipped with a nozzle that is disposed in the container, and discharges the liquid transferred from the storage tank into the container, wherein the liquid discharge section of the nozzle is disposed in the filling liquid in the middle of the filling process of the liquid.

The filling device is preferably equipped with a control device that controls the liquid discharge section of the nozzle to be in the filling liquid in the middle of the filling process of the liquid.

In the filling device of the invention, the thing that the nozzle discharging the liquid transferred from the storage tank into the container is disposed in the internal section of the container refers that the liquid discharge section of the nozzle is inserted more inward than the upper lid from the upper lid of the container.

In addition, the thing that the liquid discharge section of the nozzle is disposed in the filling liquid in the middle of the filling process refers to that the nozzle is rendered to be inserted into the liquid such that the liquid discharge section of the nozzle is in the liquid from the time when the filling liquid become accumulated in the container, although this is impossible because there is no liquid filled at the initial stage for filling. Specifically, it refers that the liquid discharge section of the nozzle is positioned in the filling liquid in the container.

Since filling of the liquid into the container is performed in such state, air inclusion at the time when the liquid is discharged from the nozzle, is suppressed. From this, the foaming is suppressed, and the filling efficiency per volume of the container improves.

The filling device of the invention is preferably equipped with a detection means that detects the status of the filling of the liquid in the container, and a flow rate-controlling means that controls the flow rate of the liquid discharged from the nozzle depending on the status of the filling of the liquid in the container, in addition to the nozzle.

Examples of the status of the filling in the container include the filling amount (weight of the liquid filled), the filling rate (the ratio of the volume of the liquid filled with respect to the internal volume of the container), or the filling step and the like.

Examples of the detection means that detects the status of the filling of the liquid include means of detecting the status of the filling using weight measurement and filling rate calculated by conversion by a liquid surface sensor, and transmitting it to the control device as a signal. Specifically, examples of the detection means include a device for weight measurement, a liquid surface sensor, or a flow rate sensor, and the like.

The control device is a device treating the signal transmitted from the detection means, and transmitting the treated signal to the flow rate-controlling means. The signal treated by the control device is transmitted to the flow rate-controlling means, and operates the flow rate-controlling means as necessary, and controls the flow rate of the liquid. The control device is preferably a device controlled by a computer.

The flow rate control is performed by controlling the flow rate of the liquid depending on the status of the filling of the liquid in the container by opening degree of the flow rate-controlling valve established in the channel of the liquid such as a pipe or a nozzle. The flow rate-controlling valve can be arranged on a pipe connecting the storage tank and the nozzle for the filling, or on the internal section of the nozzle or the discharge section of the nozzle.

Examples of the flow rate-controlling means include flow rate-controlling valves, valves, or flow rate-controlling of a pump, and the like.

The nozzle in the filling device of the invention is preferably a vertically movable nozzle. At this time, the filling device is preferably further equipped with a nozzle position-controlling means that controls the position of the tip of the movable nozzle depending on the status of the filling of the liquid in the container.

The vertically movable nozzle is a nozzle that can move vertically in the direction perpendicular to the bottom surface of the container.

As described above, the signal transmitted to the control device by the detection means is treated by the control device. When the nozzle is a vertically movable nozzle, the signal treated by the control device is transmitted to the nozzle position-controlling means, and operates the nozzle position-controlling means as necessary, and controls the position of the tip of the nozzle. The control device is preferably a device controlled by a computer.

The nozzle position-controlling means may be in any form if it can control the position of the tip of the nozzle (the height from the bottom surface to the tip of the nozzle) depending on the status of the filling of the liquid in the container.

Examples of the nozzle position-controlling means include nozzle-elevating devices, nozzle-extension devices and the like.

The filling device of the invention is preferably constituted such that the storage tank is positioned upward from the filling device, and the liquid is filled into the container by the gravity.

The thing that the storage tank is positioned upward from the filling device, and the liquid is filled into the container by the gravity refers that the liquid surface of the liquid containing at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom stored in a storage tank is always positioned upward from the filling device, namely in higher position than the upper lid of the container placed in the filling device, and the liquid in the storage tank is transferred and filled into the container by the gravity and the opening degree of the flow rate regulation valve without using a pump.

The filling device according to one aspect of the invention is preferably equipped with

a storage tank storing a liquid containing at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom;

a container for transporting or storing the liquid;

a nozzle that discharges the liquid transferred from the storage tank into the container;

a detection means that detects the status of the filling of the liquid in the container; and

a flow rate-controlling means that controls the flow rate of the liquid discharged from the nozzle depending on the status of the filling of the liquid in the container.

The filling device according to another aspect of the invention is preferably equipped with

a storage tank storing a liquid containing at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom;

a container for transporting or storing the liquid;

a vertically movable nozzle that discharges the liquid transferred from the storage tank into the container;

a detection means that detects the status of the filling of the liquid in the container; and

a nozzle position-controlling means that controls the position of the tip of the movable nozzle depending on the status of the filling of the liquid in the container.

The filling method of the invention is a method using the filling device, wherein filling of the liquid into the container is performed such that the liquid discharge section of the nozzle is positioned in the filling liquid in the middle of the filling process of the liquid. The filling method of the invention specifically has a filling step in which the tip of the nozzle is positioned in the container, and the liquid is discharged from the tip.

The filling method of the invention is suitably performed using the filling device described above, which is equipped with the nozzle, the detection means, and the flow rate-controlling means.

In the filling step, at least either or both of the flow rate at the initial stage for filling and the flow rate at the later stage for filling are preferably less than the maximum flow rate.

The initial stage for filling represents a step in which the filling rate into the container is less than 12.5%, and the later stage for filling represents a step in which the filling rate is equal to or more than 97.5%. The step between the initial stage for filling and the later stage for filling is referred to as the middle stage for filling, and the flow rate of the middle stage for filling is the maximum flow rate.

The flow rate is a volume per unit time of the liquid discharged from the nozzle, and measured with a flowmeter, a weight measurement device and the like.

If the flow rate of the liquid at the initial stage for filling, namely in the state where the liquid is little in the container, is the maximum flow rate, the linear speed of the discharge is rapid, and bubbles are easily formed by contact of the liquid with the internal wall of the container, or air inclusion, and the like. By the flow rate of the liquid at the initial stage for filling being less than the maximum flow rate, it is possible to suppress bubbles of the liquid, and enhance the filling efficiency per volume of the container.

In addition, by the flow rate at the later stage for filling being less than the maximum flow rate, it is possible to easily adjust the filling amount when the filling is completed, and enhance the filling efficiency per volume of the container.

The setting values of the filling rates at the initial stage, the middle stage, or the later stage are not limited to the setting values described above, but are preferably optimized by the bottom surface area and the height of the container to be filled, and viscosity of a bacterial liquid to be filled and easiness of bubbles.

The maximum flow rate of the liquid (the flow rate at the middle stage for filling) is set up such that the linear speed of the liquid discharge at the valve of the tip of the movable nozzle is preferably 0.2 to 3 m/second, and more preferably 0.2 to 2 m/second.

The linear speed of the discharge is a value obtained by division of the discharge flow rate by the cross-sectional area of the nozzle discharge port, and is measured with a flowmeter, or a weight measurement device and the like.

The flow rate at the initial stage for filling is desirably less than the maximum flow rate, but is preferably equal to or less than a half of the maximum flow rate, and particularly more preferably equal to or less than one fifth of the maximum flow rate from the point of excellence in the effect of improving the filling efficiency.

The lower limit is not particularly limited, but is preferably equal to or more than one hundredth of the maximum flow rate in consideration of the time required for the filling.

Specifically, the range of the flow rate at the initial stage for filling is preferably equal to or more than one hundredth and equal to or less than a half of the maximum flow rate, and more preferably equal to or more than one hundredth and is equal to or less than one fifth.

The flow rate at the later stage for filling is desirably less than the maximum flow rate, but is preferably equal to or less than a half of the maximum flow rate, and particularly more preferably equal to or less than one fifth of the maximum flow rate from the point of excellence in the bubble suppression effects. The lower limit is not particularly limited, but is preferably equal to or more than one hundredth of the maximum flow rate in consideration of the time required for the filling.

Specifically, the range of the flow rate at the later stage for filling is preferably equal to or more than one hundredth and equal to or less than a half, and more preferably equal to or more than one hundredth and equal to or less than one fifth of the maximum flow rate.

In the filling method of the invention, using a filling device, which is equipped with a vertically movable nozzle as a nozzle, and further equipped with a nozzle position-controlling means that controls the position of the tip of the movable nozzle depending on the status of the filling of the liquid in the container, the tip of the movable nozzle is preferably positioned in the proximity of the bottom surface of the container at the initial stage for filling of the filling step, and thereafter, the movable nozzle is made to rise depending on the status of the filling of the liquid in the container.

The proximity of the bottom surface of the container refers to 20% by height of the container height from the bottom surface.

The thing that the movable nozzle is made to rise depending on the status of the filling of the liquid refers that the movable nozzle is made to be inserted into the liquid so that the liquid discharge section of the movable nozzle is in the filling liquid from the time when the filling liquid is accumulated in the container. By making the liquid discharge section of the movable nozzle be present in the filling liquid, inclusion of air at the time of the liquid discharge from the movable nozzle is suppressed. From this, the foaming is suppressed, and the filling efficiency per volume of the container improves.

The length from the liquid surface of the filling liquid to the discharge section when the movable nozzle is inserted into the filling liquid is preferably 1 to 50 cm.

In addition, the liquid discharge section of the movable nozzle is made to rise in accordance to the rise of the liquid surface, whereby to suppress the volume of the nozzle soaked in the filling liquid, leading to improvement of the filling efficiency.

The volume of the nozzle soaked in the filling liquid is preferably 0.00002 to 0.005 m³.

As explained above, according to the filling device and the filling method of the invention, it is possible to suppress the foaming at the time of the filling, and improve the filling efficiency per volume of the container by controlling at least either or both of the flow rate and the position of the tip of the nozzle depending on the status of the filling of the liquid in the container at the time of the filling.

EXAMPLES

Hereinafter, the invention will be explained more specifically with Examples and Comparative Example. However, the invention is not limited thereto with respect to the intended scope.

Example 1 (1) With Respect to Embodiment

FIG. 1 is a schematic drawing that illustrates the constitution of a filling device 10 used in this Example.

The filling device 10 is equipped with

a pipe 1 of which the upstream terminal is connected to a storage tank 20 storing a liquid L containing at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom (suspension of the bacterial cells);

a vertically movable nozzle 3 arranged at the downstream terminal section of the pipe 1;

a valve 2 arranged at the tip of a movable nozzle;

a nozzle elevating device 4 that vertically migrates the movable nozzle 3;

a weight measurement device 5 that measures the weight of a container 30; and

a control device 6 that is electrically connected to the valve 2, the nozzle elevating device 4 and the weight measurement device 5.

The container 30 is placed on the weight measurement device 5, and the storage tank 20 is established upward thereof.

Specifically, as the valve 2, a valve having a lid on the tip of the nozzle, and allowing flow rate regulation with narrow or fully opening of the lid was used. By this valve 2 and by the gravity, the liquid L in the storage tank 20 can be filled into the container 30.

As the movable nozzle 3, a following-type long nozzle form was used.

As the nozzle elevating device 4, a hydraulic elevating device was used.

As the weight measurement device 5, a platform weighing machine of load cell type was used.

(2) Preparation Method of Suspension of Bacterial Cells

Rhodococcus rhodochrous J-1 strain (FERM BP-1478) having nitrile hydratase activity was aerobically cultured with a culture medium (20° C. at pH 7.0) comprising 2 mass % of glucose, 1 mass % of urea, 0.5 mass % of peptone, 0.3 mass % of yeast extract and 0.05 mass % of cobalt chloride. After completion of the culture, the cultured bacterial cells were collected by centrifuge, and this was washed with 50 mM phosphate buffer (pH 7.0 at 20° C.). The washed bacterial cells was added with the buffer to give a suspension of the bacterial cells (10 mass % in conversion to dry bacterial cells).

(3) Filling of Suspension of Bacterial Cells into Container from Storage Tank

3000 L of the suspension of the bacterial cells (10 mass % in conversion to dry bacterial cells) was added to a storage tank 20 made of stainless steel having 9 m³ of the capacity, and filling into a container 30 was initiated. The filling was performed at 0.5 L/minute by narrow-opening the valve 2 in the initial stage for filling where the filling rate was equal to or less than 12.5%, and in the later stage where the filling rate was equal to or more than 97.5%, and at 20 L/minute in the middle stage for filling by full-opening the valve 2.

The position of the movable nozzle 3 was controlled at the time of the filling such that the valve 2 of the tip of the movable nozzle 3 was always soaked in the suspension of the bacterial cells in a constant depth by the control device 6 depending on the weight value measured with the weight measurement device 5.

As a result thereof, the suspension of the bacterial cells could be filled into the container 30 while suppressing the foaming.

As described above, the foaming was suppressed when the suspension of the bacterial cells was filled, and the volume of the movable nozzle 3 soaked in the filled suspension of the bacterial cells was suppressed to the minimum, and thus the filling efficiency was improved.

INDUSTRIAL APPLICABILITY

According to the invention, it is possible to provide a filling device and a filling method, which makes it possible to effectively fill a liquid containing at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom into a container. Accordingly, the invention is useful in use and the like of a biocatalyst in the industrial process field and the like.

EXPLANATIONS OF LETTERS OR NUMERALS

1: Pipe

2: Valve

3: Movable nozzle

4: Nozzle elevating device

5: Weight measurement device

6: Control device

10: Filling device

20: Storage tank

30: Container

L: Liquid 

1. A filling device for filling a liquid containing at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom stored in a storage tank into a container, which is equipped with a nozzle that is disposed in a container, and discharges the liquid transferred from the storage tank into the container, wherein the liquid discharge section of the nozzle is disposed in the filling liquid in the middle of the filling process of the liquid.
 2. A filling device for filling a liquid containing at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom stored in a storage tank into a container, which is equipped with a nozzle that discharges the liquid transferred from the storage tank into the container; a detection means that detects the status of the filling of the liquid in the container; and a flow rate-controlling means that controls the flow rate of the liquid discharged from the nozzle depending on the status of the filling of the liquid in the container.
 3. A filling device for filling a liquid containing at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom stored in a storage tank into a container, which is equipped with a vertically movable nozzle that discharges the liquid transferred from the storage tank into the container; a detection means that detects the status of the filling of the liquid in the container; and a nozzle position-controlling means that controls the position of the tip of the movable nozzle depending on the status of the filling of the liquid in the container.
 4. The filling device according to claim 1, which is constituted such that the storage tank is positioned upward from the filling device, and the liquid is filled into the container by the gravity.
 5. The filling device according to claim 1, wherein at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom have nitrile hydratase activity.
 6. A filling method for filling a liquid containing at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom stored in a storage tank into a container, the filling method comprising a filling step in which the tip of a nozzle of a filling device is positioned in the container, and the liquid is discharged from the tip, wherein the filling device is equipped with a nozzle that discharges the liquid transferred from the storage tank into the container; a detection means that detects the status of the filling of the liquid in the container; and a flow rate-controlling means that controls the flow rate of the liquid discharged from the nozzle depending on the status of the filling of the liquid in the container, and wherein in the filling step, at least either or both of the flow rate at the initial stage for filling and the flow rate at the later stage for filling are less than the maximum flow rate.
 7. The filling method according to claim 6, wherein the flow rate at the initial stage for filling of the filling step is equal to or less than a half of the maximum flow rate.
 8. The filling method according to claim 6, wherein the flow rate at the later stage for filling of the filling step is equal to or less than a half of the maximum flow rate.
 9. A filling method for filling a liquid containing at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom stored in a storage tank into a container, the filling method comprising a filling step in which the tip of a movable nozzle of a filling device is positioned in the container, and the liquid is discharged from the tip, wherein the filling device is equipped with a vertically movable nozzle that discharges the liquid transferred from the storage tank into the container; a detection means that detects the status of the filling of the liquid in the container; and the nozzle position-controlling means that controls the position of the tip of the movable nozzle depending on the status of the filling of the liquid in the container, and wherein the tip of the movable nozzle is positioned in the proximity of the bottom surface of the container at the initial stage for filling of the filling step, and thereafter, the movable nozzle is made to rise depending on the status of the filling of the liquid in the container.
 10. The filling method according to claim 6, wherein at least either or both of the bacterial cells of a microorganism and the treatment substance obtained therefrom have nitrile hydratase activity.
 11. The filling device according to claim 2, which is constituted such that the storage tank is positioned upward from the filling device, and the liquid is filled into the container by the gravity.
 12. The filling device according to claim 3, which is constituted such that the storage tank is positioned upward from the filling device, and the liquid is filled into the container by the gravity.
 13. The filling device according to claim 2, wherein at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom have nitrile hydratase activity.
 14. The filling device according to claim 3, wherein at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom have nitrile hydratase activity.
 15. The filling device according to claim 4, wherein at least either or both of bacterial cells of a microorganism and a treatment substance obtained therefrom have nitrile hydratase activity.
 16. The filling method according to claim 7, wherein the flow rate at the later stage for filling of the filling step is equal to or less than a half of the maximum flow rate.
 17. The filling method according to claim 7, wherein at least either or both of the bacterial cells of a microorganism and the treatment substance obtained therefrom have nitrile hydratase activity.
 18. The filling method according to claim 8, wherein at least either or both of the bacterial cells of a microorganism and the treatment substance obtained therefrom have nitrile hydratase activity.
 19. The filling method according to claim 16, wherein at least either or both of the bacterial cells of a microorganism and the treatment substance obtained therefrom have nitrile hydratase activity.
 20. The filling method according to claim 9, wherein at least either or both of the bacterial cells of a microorganism and the treatment substance obtained therefrom have nitrile hydratase activity. 